Bridged bicyclic amine derivatives useful as CCR-3 receptor antagonists

ABSTRACT

Compounds having the formula (1),  
                 
 
are useful as CCR-3 receptor antagonists, wherein T is a bridged heterocyclyl group having one N atom and a bridge of one to two bridgehead carbon atoms; Ar and Ar 1  are aryl or heteroaryl; F is alkylene, alkenylene, or a bond; E is —C(═O)N(R 10 )—, —SO 2 N(R 10 )—, —N(R 11 )C(═O)N(R 10 )—, —N(R 11 )SO 2 N(R 10 )—, —N(R 11 )C(═S)N(R 10 )—, —N(R 11 )C(═O)—, —N(R 11 )SO 2 —, —N(R 12 )C(═O)CH(R 13 )—, or CH(R 13 )C(═O)N(R 12 )—; Q is —C(═O)— or C 1-2 alkylene; and R 3 , R 4 , R 5 , R 9 , R 10 , R 11 , R 12 , and R 13  are defined as set forth in the specification.

PRIORITY

This application claims priority from U.S. Ser. No. 60/450,380, filed 27Feb. 2003, incorporated herein by reference in full.

FIELD OF THE INVENTION

The invention relates to certain bridged bicyclic amine derivatives thatare CCR-3 receptor antagonists, as well as pharmaceutical compositionscontaining them and methods for their use.

BACKGROUND INFORMATION

Tissue eosinophilia is a feature of a number of pathological conditionssuch as asthma, rhinitis, eczema and parasitic infections (see Bousquet,J. et al., N. Eng. J. Med. 323:1033-39 (1990) and Kay, A. B. andCorrigan, C. J., Br. Med. Bull. 48:51-64 (1992)). In asthma, eosinophilaccumulation and activation are associated with damage to bronchialepithelium and hyperresponsiveness to constrictor mediators. Chemokinessuch as RANTES, eotaxin and MCP-3 are known to activate eosinophils (seeBaggiolini, M. and Dahinden, C. A., Immunol. Today, 15:127-33 (1994),Rot, A. M. et al., J. Exp. Med. 176:1489-95 (1992) and Ponath, P. D. etal., J. Clin. Invest., 97(3):604-12 (1996)). However, unlike RANTES andMCP-3 which also induce the migration of other leukocyte cell types,eotaxin is selectively chemotactic for eosinophils (seeGriffith-Johnson, D. A. et al., Biochem. Biophy. Res. Commun. 197:1167(1993), and Jose, P. J. et al., Biochem. Biophy. Res. Commun. 207:788(1994)). Specific eosinophil accumulation has been observed at the siteof administration of eotaxin, whether by intradermal or intraperitonealinjection or aerosol inhalation (see Griffith-Johnson, D. A. et al.,supra; Jose, P. J. et al., supra; Rothenberg, M. E. et al., J. Exp. Med.181:1211 (1995), and Ponath, P. D., supra).

Glucocorticoids such as dexamethasone, methprednisolone andhydrocortisone have been used for treating many eosinophil-relateddisorders, including bronchial asthma (R. P. Schleimer et al., Am. Rev.Respir. Dis., 141:559 (1990)). The glucocorticoids are believed toinhibit IL-5 and IL-3 mediated eosinophil survival in these diseases.However, prolonged use of glucocorticoids can lead to side effects inpatients such as glaucoma, osteoporosis, and growth retardation (seeHanania, N. A. et al., J. Allergy and Clin. Immunol., 96:571-79 (1995)and Saha, M. T. et al., Acta Paediatrica, 86(2):138-42 (1997)). It istherefore desirable to have an alternative means of treatingeosinophil-related diseases without incurring these undesirable sideeffects.

Recently, the CCR-3 receptor was identified as a major chemokinereceptor that eosinophils use for their response to eotaxin, RANTES andMCP-3. When transfected into a murine pre-beta lymphoma line, CCR-3bound eotaxin, RANTES and MCP-3 conferred chemotactic responses on thesecells to eotaxin, RANTES and MCP-3 (see Ponath, P. D. et al., J. Exp.Med., 183:2437-48 (1996)). The CCR-3 receptor is expressed on thesurface of eosinophils, T-cells (subtype Th-2), basophils and mast cellsand is highly selective for eotaxin. Studies have shown thatpretreatment of eosinophils with an anti-CCR-3 mAb completely inhibitseosinophil chemotaxis to eotaxin, RANTES and MCP-3 (see Heath, H. etal., J. Clin. Invest., 99(2):178-84 (1997)). U.S. Pat. Nos. 6,140,344and 6,166,015 issued to Applicant herein and EP application EP903349,published Mar. 24, 1999 disclose CCR-3 antagonists that inhibiteosinophilic recruitment by chemokine such as eotaxin.

SUMMARY OF THE INVENTION

The present invention is directed to bridged cyclic amine derivativesuseful as CCR3 receptor antagonists which are capable of inhibiting thebinding of eotaxin to the CCR-3 receptor and thereby provide a means ofcombating eosinophil induced diseases, such as asthma.

In a first aspect, this invention provides a compound of Formula (I):

wherein:

T is

where R⁶ is taken together with one of R⁷ and R⁸ to form a bridge of oneto two bridgehead carbon atoms, and the other of R⁷ and R⁸ is selectedfrom hydrogen and R⁹;

Ar and Ar¹ are, independently of each other, aryl or heteroaryl;

F is alkylene, alkenylene, or a bond;

-   -   E is selected from —C(═O)N(R¹⁰)—, —SO₂N(R¹⁰)—,        —N(R¹¹)C(═O)N(R¹⁰)—, —N(R¹¹)SO₂N(R¹⁰)—, —N(R¹¹)C(═S)N(R¹⁰)—,        —N(R¹¹)C(═O)—, —N(R¹¹)SO₂—, —N(R¹²)C(═O)CH(R¹³)—, and        CH(R¹³)C(═O)N(R¹²)—, where:        -   R¹⁰, R¹¹, R¹², and R¹³ are, independently of each other,            hydrogen, alkyl, acyl, haloalkyl, cycloalkyl,            cycloalkylalkyl, aryl, aralkyl, aralkenyl, heteroaryl,            heteroaralkyl, heterocycloalkyl, heteroalkyl, or            -(alkylene)-C(═O)-Z, where Z is alkyl, haloalkyl, alkoxy,            haloalkyloxy, hydroxy, amino, mono- or disubstituted amino,            aryl, aralkyl, aryloxy, aralkyloxy, heteroaryl,            heteroaryloxy, or heteroaralkyloxy;        -   or alternatively, R¹² and R¹³ may be taken together with the            nitrogen and carbon atoms to which they are attached,            respectively, to form a heterocyclyl or heteroaryl ring            optionally substituted with up to two groups selected from            R¹⁴;    -   R³ and R⁴ are, independently of each other, hydrogen, alkyl,        alkenyl, haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl,        heteroaryl, heteroaralkyl, heterocyclyl, heterocyclylalkyl,        heteroalkyl, -(alkylene)-C(═O)-Z¹, or -(alkylene)-C(═O)₂Z¹,        where Z¹ is alkyl, haloalkyl, alkoxy, haloalkyloxy, hydroxy,        amino, mono- or disubstituted amino, aryl, aralkyl, aryloxy,        aralkyloxy, heteroaryl, heteroaryloxy, or heteroaralkyloxy;

R⁵ is hydrogen or alkyl;

Q is —C(═O)— or C₁₋₂alkylene;

R⁹ is attached to any available carbon atom of ring T and is selectedfrom lower alkyl, hydroxy, lower alkoxy, halo, cyano, trifluoromethyl,trifluoromethoxy, or a lower alkyl substituted with one of hydroxy,lower alkoxy, halo, cyano, trifluoromethyl, or trifluoromethoxy;

R¹⁴ is selected from lower alkyl, hydroxy, lower alkoxy, halo, cyano,trifluoromethyl, trifluoromethoxy, and a lower alkyl substituted withone of hydroxy, lower alkoxy, halo, cyano, trifluoromethyl, ortrifluoromethoxy;

m is 0 or 1; and

n is 0 to 4; and

prodrugs, individual isomers, mixtures of isomers, and pharmaceuticallyacceptable salts thereof.

The invention also relates to pharmaceutical compositions containingcompounds of Formula (I), above, and methods of treating CCR-3 receptormediated diseases, such as asthma, by administration of atherapeutically-effective amount of a compound of Formula (I), to apatient in need of treatment thereof.

DETAILED DESCRIPTION OF THE INVENTION

Definitions

Unless otherwise stated, the following terms used in the specificationand claims have the meanings given below.

“Alkyl” means a linear saturated monovalent hydrocarbon radical of oneto six carbon atoms or a branched saturated monovalent hydrocarbonradical of three to six carbon atoms, e.g., methyl, ethyl, propyl,2-propyl, n-butyl, iso-butyl, tert-butyl, pentyl, and the like. A “loweralkyl” is an alkyl group having one to four carbon atoms.

“Alkenyl” means a linear monovalent hydrocarbon radical of two to sixcarbon atoms or a branched monovalent hydrocarbon radical of three tosix carbon atoms, containing at least one double bond, e.g., ethenyl,propenyl, and the like.

When the term “alkyl” is used as a suffix following another term, as in“phenylalkyl,” or “hydroxyalkyl,” this is intended to refer to an alkylgroup, as defined above, being substituted with at least one substituentselected from the other specifically named group. Thus, “phenylalkyl”would include, for example, benzyl and phenylethyl. “Hydroxyalkyl”includes, for example, 2-hydroxyethyl, 2-hydroxypropyl, 3-hydroxypropyl,1-(hydroxymethyl)-2-methylpropyl, 2-hydroxybutyl,2-hydroxy-1-hydroxymethylethyl, 2,3-dihydroxybutyl, 3,4-dihydroxybutyland 2-(hydroxymethyl)-3-hydroxypropyl, and so forth. Accordingly, asused herein, the term “hydroxyalkyl” is used to define a subset ofheteroalkyl groups defined below.

“Acyl” means a radical —C(═O)R, where R is hydrogen, alkyl, cycloalkyl,cycloalkylalkyl, phenyl or phenylalkyl wherein the alkyl, cycloalkyl,cycloalkylalkyl, and phenylalkyl groups are as defined herein.Representative examples include, but are not limited to formyl, acetyl,cyclohexylcarbonyl, cyclohexylmethylcarbonyl, benzoyl, benzylcarbonyl,and the like.

“Acylamino” means a radical —NR′C(═O)R, where R′ is hydrogen or alkyl,and R is hydrogen, alkyl, cycloalkyl, cycloalkylalkyl, phenyl orphenylalkyl, wherein the alkyl, cycloalkyl, cycloalkylalkyl, andphenylalkyl groups are as defined herein. Representative examplesinclude, but are not limited to formylamino, acetylamino,cylcohexylcarbonylamino, cyclohexylmethyl-carbonylamino, benzoylamino,benzylcarbonylamino, and the like.

“Alkoxy” means a radical —OR where R is an alkyl as defined herein e.g.,methoxy, ethoxy, propoxy, butoxy and the like. A “lower alkoxy” is analkoxy group wherein the alkyl (R) group has up to four carbon atoms.

“Alkoxycarbonyl” means a radical —C(═O)R where R is alkoxy is as definedherein.

“Alkylamino” or “Monoalkylamino” means a radical —NHR where R representsan alkyl, cycloalkyl or cycloalkyl-alkyl group as defined herein.Representative examples include, but are not limited to methylamino,ethylamino, isopropylamino, cyclohexylamino, and the like.

“Alkylene” means a linear saturated bivalent hydrocarbon radical of oneto six carbon atoms or a branched saturated bivalent hydrocarbon radicalof three to six carbon atoms, e.g., methylene, ethylene,2,2-dimethylethylene, propylene, 2-methylpropylene, butylene, pentylene,and the like.

“Alkynyl” means a linear monovalent hydrocarbon radical of two to sixcarbon atoms or a branched monovalent hydrocarbon radical of three tosix carbon atoms, containing at least one triple bond, e.g., ethynyl,propynyl, and the like.

“Alkylsulfonyl” means a radical —S(O)₂R where R is an alkyl, cycloalkylor cycloalkyl-alkyl group as defined herein, e.g., methylsulfonyl,ethylsulfonyl, propylsulfonyl, butylsulfonyl, cyclohexylsulfonyl and thelike.

“Alkylsulfinyl” means a radical —S(O)R where R is an alkyl, cycloalkylor cycloalkyl-alkyl group as defined herein e.g., methylsulfinyl,ethylsulfinyl, propylsulfinyl, butylsulfinyl, cyclohexylsulfonyl and thelike.

“Alkylthio” means a radical —SR where R is an alkyl as defined abovee.g., methylthio, ethylthio, propylthio, butylthio, and the like.

“Aryl” means a monocyclic or bicyclic aromatic hydrocarbon radical whichis optionally substituted with one or more substituents, preferably one,two or three, substituents selected from the group consisting of alkyl,haloalkyl, hydroxyalkyl, heteroalkyl, acyl, acylamino, amino,alkylamino, dialkylamino, alkylthio, alkylsulfinyl, alkylsulfonyl,—SO₂NR′R″ (where R′ and R″ are independently hydrogen or alkyl), alkoxy,haloalkoxy, alkoxycarbonyl, carbamoyl, hydroxy, halo, nitro, cyano,mercapto, methylenedioxy or ethylenedioxy. Exemplary aryl groupsinclude, but are not limited to, phenyl, chlorophenyl, fluorophenyl,methoxyphenyl, 1-naphthyl, 2-naphthyl, and the derivatives thereof.

“Cycloalkyl” refers to a saturated monovalent cyclic hydrocarbon radicalof three to seven ring carbons e.g., cyclopropyl, cyclobutyl,cyclohexyl, 4-methylcyclohexyl, and the like.

“Dialkylamino” means a radical —NRR′ where R and R′ independentlyrepresent an alkyl, cycloalkyl, or cycloalkylalkyl group as definedherein. Representative examples include, but are not limited todimethylamino, methylethylamino, di(1-methylethyl)amino,(cyclohexyl)(methyl)amino, (cyclohexyl)(ethyl)amino,(cyclohexyl)(propyl)amino, (cyclohexylmethyl)(methyl)amino,(cyclohexylmethyl)(ethyl)amino, and the like.

“Halo” means fluoro, chloro, bromo, or iodo, preferably fluoro andchloro.

“Haloalkyl” means alkyl substituted with one or more same or differenthalo atoms, e.g., —CH₂Cl, —CF₃, —CH₂CF₃, —CH₂CCl₃, and the like.

“Heteroaryl” means a monocyclic or bicyclic radical of 5 to 12 ringatoms having at least one aromatic ring containing one, two, or threering heteroatoms selected from N, O, or S, the remaining ring atomsbeing C, with the understanding that when the heteroaryl group is abicyclic system in which one of the rings is carbocyclic and/ornon-aromatic, the point of attachment to the heteroaryl group will be tothe aromatic ring containing at least one heteroatom. The heteroarylring is optionally substituted independently with one or moresubstituents, preferably one or two substituents, selected from alkyl,haloalkyl, hydroxyalkyl, heteroalkyl, acyl, acylamino, amino,alkylamino, dialkylamino, alkylthio, alkylsulfinyl, alkylsulfonyl,—SO₂NR′R″ (where R′ and R″ are independently hydrogen or alkyl), alkoxy,haloalkoxy, alkoxycarbonyl, carbamoyl, hydroxy, halo, nitro, cyano,mercapto, methylenedioxy, ethylenedioxy or optionally substitutedphenyl. More specifically the term heteroaryl includes, but is notlimited to, pyridyl, furanyl, thienyl, thiazolyl, isothiazolyl,triazolyl, imidazolyl, isoxazolyl, pyrrolyl, pyrazolyl, pyrimidinyl,5-(3,4-dimethoxyphenyl)-pyrimidin-2-yl,5-(4-methoxyphenyl)-pyrimidin-2-yl,5-(3,4-methylenedioxyphenyl)-pyrimidin-2-yl, benzofuranyl,tetrahydrobenzofuranyl, isobenzofuranyl, benzothiazolyl,benzoisothiazolyl, benzotriazolyl, indolyl, isoindolyl, benzoxazolyl,quinolyl, tetrahydroquinolinyl, isoquinolyl, benzimidazolyl,benzisoxazolyl or benzothienyl and derivatives thereof.

“Heteroalkyl” means an alkyl radical as defined herein wherein one, twoor three hydrogen atoms have been replaced with a substituentindependently selected from the group consisting of —OR^(a),—NR^(b)R^(c), and —S(O)_(n)R^(d) (where n is an integer from 0 to 2),with the understanding that the point of attachment of the heteroalkylradical is through a carbon atom, wherein R^(a) is hydrogen, acyl,alkyl, cycloalkyl, or cycloalkylalkyl; R^(b) and R^(c) are independentlyof each other hydrogen, acyl, alkyl, cycloalkyl, or cycloalkylalkyl; andwhen n is 0, R^(d) is hydrogen, alkyl, cycloalkyl, or cycloalkylalkyl,and when n is 1 or 2, R^(d) is alkyl, cycloalkyl, cycloalkylalkyl,amino, acylamino, monoalkylamino, or dialkylamino. Representativeexamples include, but are not limited to, 2-hydroxyethyl,3-hydroxypropyl, 2-hydroxy-1-hydroxymethylethyl, 2,3-dihydroxypropyl,1-hydroxymethylethyl, 3-hydroxybutyl, 2,3-dihydroxybutyl,2-hydroxy-1-methylpropyl, 2-aminoethyl, 3-aminopropyl,2-methylsulfonylethyl, aminosulfonylmethyl, aminosulfonylethyl,aminosulfonylpropyl, methylaminosulfonylmethyl,methylaminosulfonylethyl, methylaminosulfonylpropyl, and the like.

“Heterocyclyl” means a saturated or unsaturated non-aromatic cyclicradical of 3 to 8 ring atoms in which one or two ring atoms areheteroatoms selected from NR^(x) {wherein each R^(x) is independentlyhydrogen, alkyl, acyl, alkylsulfonyl, aminosulfonyl,(alkylamino)sulfonyl, (dialkylamino)sulfonyl, carbamoyl,(alkylamino)carbonyl, (dialkylamino)carbonyl, (carbamoyl)alkyl,(alkylamino)carbonylalkyl, or dialkylaminocarbonylalkyl}, O, or S(O)_(n)(where n is an integer from 0 to 2), the remaining ring atoms beingcarbon atoms. The heterocyclyl ring may be optionally substitutedindependently with one, two, or three substituents selected from alkyl,haloalkyl, heteroalkyl, halo, nitro, cyanoalkyl, hydroxy, alkoxy, amino,monoalkylamino, dialkylamino, aralkyl, —(X)_(n)—C(═O)R (where X is O orNR′, n is 0 or 1, R is hydrogen, alkyl, haloalkyl, hydroxy, alkoxy,amino, monoalkylamino, dialkylamino or optionally substituted phenyl,and R′ is hydrogen or alkyl), -alkylene-C(═O)R (where R is hydrogen,alkyl, haloalkyl, hydroxy, alkoxy, amino, monoalkylamino, dialkylaminoor optionally substituted phenyl) or —S(O)_(n)R^(d) (where n is aninteger from 0 to 2, and R^(d) is hydrogen (provided that n is 0),alkyl, haloalkyl, cycloalkyl, cycloalkylalkyl, amino, monoalkylamino,dialkylamino, or hydroxyalkyl). More specifically the term heterocyclylincludes, but is not limited to, tetrahydropyranyl, piperidino,N-methylpiperidin-3-yl, piperazino, N-methylpyrrolidin-3-yl,3-pyrrolidino, morpholino, thiomorpholino, thiomorpholino-1-oxide,thiomorpholino-1,1-dioxide, tetrahydrothiophenyl-S,S-dioxide,pyrrolinyl, imidazolinyl, and the derivatives thereof.

“Leaving group” has the meaning conventionally associated with it insynthetic organic chemistry, i.e., an atom or a group capable of beingdisplaced by a nucleophile and includes halo (such as chloro, bromo, andiodo), alkanesulfonyloxy, arenesulfonyloxy, alkylcarbonyloxy (e.g.,acetoxy), arylcarbonyloxy, mesyloxy, tosyloxy,trifluoromethanesulfonyloxy, aryloxy (e.g., 2,4-dinitrophenoxy),methoxy, N,O-dimethylhydroxylamino, and the like.

“Optionally substituted phenyl” means a phenyl group which is optionallysubstituted with one or more substituents, preferably one, two or three,substituents preferably selected from the group consisting of alkyl,haloalkyl, hydroxyalkyl, heteroalkyl, acyl, acylamino, amino,alkylamino, dialkylamino, alkylthio, alkylsulfinyl, alkylsulfonyl,—SO₂NR′R″ (where R′ and R″ are independently hydrogen or alkyl), alkoxy,haloalkoxy, alkoxycarbonyl, carbamoyl, hydroxy, halo, nitro, cyano,mercapto, methylenedioxy or ethylenedioxy. More specifically the termincludes, but is not limited to, phenyl, chlorophenyl, fluorophenyl,bromophenyl, methylphenyl, ethylphenyl, methoxyphenyl, cyanophenyl,4-nitrophenyl, 4-trifluoromethylphenyl, 4-chlorophenyl,3,4-difluorophenyl, 2,3-dichlorophenyl, 3-methyl-4-nitrophenyl,3-chloro-4-methylphenyl, 3-chloro-4-fluorophenyl or 3,4-dichlorophenyland the derivatives thereof.

“Optional” or “optionally” means that the subsequently described eventor circumstance may but need not occur, and that the descriptionincludes instances where the event or circumstance occurs and instancesin which it does not. For example, “aryl group optionally mono- ordi-substituted with an alkyl group” means that the alkyl may but neednot be present, and the description includes situations where the arylgroup is mono- or disubstituted with an alkyl group and situations wherethe aryl group is not substituted with the alkyl group.

“Substituted alkyl” means an alkyl group having one or more, preferablyone, two or three substituents selected from the group consisting ofacyl, acylamino, hydroxy, alkoxy, amino, haloalkyl, halo,alkoxycarbonyl, alkylamino, alkylsulfonyl, alkylsulfinyl, alkylthio,aryl, cycloalkyl, dialkylamino, heteroaryl and/or heterocyclyl, asdefined above.

“Pharmaceutically acceptable excipient” means an excipient that isuseful in preparing a pharmaceutical composition that is generally safe,non-toxic and neither biologically nor otherwise undesirable, andincludes excipients that are acceptable for veterinary use as well ashuman pharmaceutical use. A “pharmaceutically acceptable excipient” asused in the specification and claims includes both one and more than onesuch excipient.

“Pharmaceutically acceptable salt” of a compound means a salt that ispharmaceutically acceptable and that possesses the desiredpharmacological activity of the parent compound. Such salts include: (1)acid addition salts, formed with inorganic acids such as hydrochloricacid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, andthe like; or formed with organic acids such as acetic acid, propionicacid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvicacid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid,fumaric acid, tartaric acid, citric acid, benzoic acid,3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid,methanesulfonic acid, ethanesulfonic acid, 1,2-ethane-disulfonic acid,2-hydroxyethanesulfonic acid, benzenesulfonic acid,4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid,4-toluenesulfonic acid, camphorsulfonic acid,4-methylbicyclo[2.2.2]-oct-2-ene-1-carboxylic acid, glucoheptonic acid,3-phenylpropionic acid, trimethylacetic acid, tertiary butylacetic acid,lauryl sulfuric acid, gluconic acid, glutamic acid, hydroxynaphthoicacid, salicylic acid, stearic acid, muconic acid, and the like; or (2)salts formed when an acidic proton present in the parent compound eitheris replaced by a metal ion, e.g., an alkali metal ion, an alkaline earthion, or an aluminum ion; or coordinates with an organic base such asethanolamine, diethanolamine, triethanolamine, tromethamine,N-methylglucamine, and the like.

A “prodrug” of a compound of formula (I) herein refers to any compoundwhich releases an active drug according to Formula I in vivo when suchprodrug is administered to a mammalian subject. Prodrugs of a compoundof Formula I are prepared by modifying one or more functional group(s)present in the compound of Formula I in such a way that themodification(s) may be cleaved in vivo to release the compound ofFormula I. Prodrugs include compounds of Formula I wherein a hydroxy,amino, or sulfhydryl group in a compound of Formula I is bonded to anygroup that may be cleaved in vivo to regenerate the free hydroxyl,amino, or sulfhydryl group, respectively. Examples of prodrugs include,but are not limited to, esters (e.g., acetate, formate, and benzoatederivatives), carbamates (e.g., N,N-dimethylaminocarbonyl) of hydroxyfunctional groups in compounds of Formula I, and the like.

“Protecting group” refers to a grouping of atoms that when attached to areactive group in a molecule masks, reduces or prevents that reactivity.Examples of protecting groups can be found in T. W. Green and P. G.Wuts, Protective Groups in Organic Chemistry, (Wiley, 2^(nd) ed. 1991)and Harrison and Harrison et al., Compendium of Synthetic OrganicMethods, Vols. 1-8 (John Wiley and Sons, 1971-1996). Representativeamino protecting groups include, formyl, acetyl, trifluoroacetyl,benzyl, benzyloxycarbonyl (CBZ), tert-butoxycarbonyl (Boc), trimethylsilyl (TMS), 2-trimethylsilylethanesulfonyl (SES), trityl andsubstituted trityl groups, allyloxycarbonyl,9-fluorenylmethyloxycarbonyl (FMOC), nitro-veratryloxycarbonyl (NVOC),and the like. Representative hydroxy protecting groups include thosewhere the hydroxy group is either acylated or alkylated such as benzyl,and trityl ethers as well as alkyl ethers, tetrahydropyranyl ethers,trialkylsilyl ethers and allyl ethers.

“Treating” or “treatment” of a disease includes: (1) preventing thedisease, i.e., causing the clinical symptoms of the disease not todevelop in a mammal that may be exposed to or predisposed to the diseasebut does not yet experience or display symptoms of the disease; (2)inhibiting the disease, i.e., arresting or reducing the development ofthe disease or its clinical symptoms; or (3) relieving the disease,i.e., causing regression of the disease or its clinical symptoms.

“A therapeutically effective amount” means the amount of a compoundthat, when administered to a mammal for treating a disease, issufficient to effect such treatment for the disease. The“therapeutically effective amount” will vary depending on the compound,the disease and its severity and the age, weight, etc., of the mammal tobe treated.

Compounds that have the same molecular formula but differ in the natureor sequence of bonding of their atoms or the arrangement of their atomsin space are termed “isomers.” Isomers that differ in the arrangement oftheir atoms in space are termed “stereoisomers”. Stereoisomers that arenot mirror images of one another are termed “diastereomers” and thosethat are non-superimposable mirror images of each other are termed“enantiomers”. When a compound has an asymmetric center, for example, ifa carbon atom is bonded to four different groups, a pair of enantiomersis possible. An enantiomer can be characterized by the absoluteconfiguration of its asymmetric center and is described by the R- andS-sequencing rules of Cahn and Prelog, or by the manner in which themolecule rotates the plane of polarized light and designated asdextrorotatory or levorotatory (i.e., as (+) or (−)-isomersrespectively). A chiral compound can exist as either individualenantiomer or as a mixture thereof. A mixture containing equalproportions of the enantiomers is called a “racemic mixture.”

The compounds of this invention may possess one or more asymmetriccenters; such compounds can therefore be produced as individual (R)- or(S)-stereoisomers or as mixtures thereof. Unless indicated otherwise,the description or naming of a particular compound in the specificationand claims is intended to include both individual enantiomers andmixtures, racemic or otherwise, thereof. The methods for thedetermination of stereochemistry and the separation of stereoisomers arewell-known in the art (see discussion in Chapter 4 of “Advanced OrganicChemistry”, 4th edition J. March, John Wiley and Sons, New York, 1992).

Preferred Embodiments

While the broadest definition of this invention is set forth in theSummary of the Invention, certain compounds of Formula (I) arepreferred.

Preferred compounds of the invention are compounds having Formula (I):

wherein:

T is

where R⁶ is taken together with one of R⁷ and R⁸ to form a bridge of oneto two bridgehead carbon atoms, and the other of R⁷ and R⁸ is selectedfrom hydrogen and R⁹;

Ar and Ar¹ are both phenyl;

F is a bond;

-   -   E is selected from —C(═O)N(R¹⁰)—, —N(R¹¹)C(═O)N(R¹⁰)—,        —N(R¹¹)C(═O)—, —N(R¹²)C(═O)CH(R¹³)—, and CH(R¹³)C(═O)N(R¹²)—,        where:        -   R¹⁰, R¹¹, R¹², and R¹³ are, independently of each other,            hydrogen or alkyl;        -   or alternatively, R¹² and R¹³ may be taken together with the            nitrogen and carbon atoms to which they are attached,            respectively, to form a heterocyclyl or heteroaryl ring            optionally substituted with up to two groups selected from            R¹⁴;    -   R³ and R⁴ are, independently of each other, hydrogen, alkyl, or        substituted alkyl (more preferably lower alkyl optionally        substituted with hydroxy);

R⁵ is hydrogen or alkyl;

Q is —C(═O)— or C₁₋₂alkylene;

R⁹ is attached to any available carbon atom of ring T and is selectedfrom lower alkyl, hydroxy, lower alkoxy, halo, cyano, trifluoromethyl,trifluoromethoxy, or a lower alkyl substituted with one of hydroxy,lower alkoxy, halo, cyano, trifluoromethyl, or trifluoromethoxy;

R¹⁴ is selected from lower alkyl, hydroxy, lower alkoxy, halo, cyano,trifluoromethyl, trifluoromethoxy, and a lower alkyl substituted withone of hydroxy, lower alkoxy, halo, cyano, trifluoromethyl, ortrifluoromethoxy;

m is 0 or 1; and

n is 0 to 4; and

prodrugs, individual isomers, mixtures of isomers, and pharmaceuticallyacceptable salts thereof.

Accordingly, in compounds of Formula (I), Ar and Ar¹ are preferably bothphenyl, more preferably phenyl optionally substituted with one, two, orthree groups selected from halo, alkyl, heteroalkyl, alkoxy, nitro,trifluoromethyl, and alkylsulfonyl. More preferably, Ar is selected fromphenyl, 4-chlorophenyl, 4-methylphenyl, 4-methoxyphenyl,3-methylsulfonylphenyl, 3,5-dimethoxyphenyl, 3,4-dimethoxyphenyl, and3,4,5-trimethoxyphenyl, and Ar¹ is preferably selected from4-chlorophenyl and 3,4-dichlorophenyl. Most preferred are compoundswhere Ar is 3,4,5-trimethoxyphenyl and Ar¹ is 4-chlorophenyl.

In compounds of Formula (I), F is preferably a bond and Q is CH₂—.

In compounds of formula (I), E is preferably selected from—C(═O)N(R¹⁰)—, —N(R¹¹)C(═O)N(R¹⁰)—, and —N(R¹²)C(═O)CH(R¹³)—, where R¹⁰,R¹¹, R¹² , and R¹³ are, independently of each other, hydrogen or alkyl;or alternatively, R¹² and R¹³ may be taken together with the nitrogenand carbon atoms to which they are attached, respectively, to form

where R¹⁸ and R¹⁹ are selected from hydrogen and lower alkyl. Incompounds of formula (I), preferably when E is

R¹⁸ and R¹⁹ are preferably methyl, and m is preferably 0. Most preferredare compounds where E is —NHC(═O)NH—.

In compounds of Formula (I), preferably T is selected from the groupconsisting of:

wherein R⁹ is attached to any available carbon atom of ring T and isselected from lower alkyl and hydroxy, and n is 0 to 2. More preferablyT is

In compounds of Formula (I), preferably R³ is hydrogen; and R⁴ ishydrogen or optionally substituted lower alkyl; more preferably R³ ishydrogen and R⁴ is methyl, ethyl, 1-methylethyl, isopropyl,1-hydroxyethyl or 2-hydroxyethyl. Further preferred compounds are thosewherein R³ is hydrogen; and R⁴ is 1-methylethyl or 1-hydroxyethyl.

According to another aspect of the invention, preferred compounds arethose compounds having the formula (II):

and pharmaceutically-acceptable salts thereof, in which:

T is

where R⁶ is taken together with one of R⁷ and R⁸ to form a bridge of oneto two bridgehead carbon atoms optionally substituted with one to twoCH₃, and the other of R⁷ and R⁸ is selected from hydrogen and loweralkyl;

-   -   E is selected from —C(═O)N(R¹⁰)—, —N(R¹¹)C(═O)N(R¹⁰)—, and        —N(R¹²)C(═O)CH(R¹³)—, where:        -   R¹⁰, R¹¹, R¹², and R¹³ are independently of each other            hydrogen or lower alkyl, or alternatively, R¹² and R¹³ may            be taken together with the nitrogen and carbon atoms to            which they are attached, respectively, to form a            five-membered heterocyclyl or heteroaryl ring having up to            two N atoms and optionally substituted with up to two groups            selected from methyl, ethyl, hydroxy, methoxy, halo, cyano,            trifluoromethyl, and trifluoromethoxy;

R⁴ is hydrogen, lower alkyl, or lower alkyl substituted with hydroxy;

R²⁰ and R²¹ are each independently selected from halo, OR²², and SO₂R²²,wherein R²² is lower alkyl;

m is 0 or 1;

p and q are independently 0, 1, 2 or 3.

In compounds of formula (II), preferably E is selected from —C(═O)NH—,—NHC(═O)NH—, and

where R¹⁸ and R¹⁹ are each hydrogen or lower alkyl; preferably R⁴ ishydrogen, methyl, ethyl, 1-hydroxyethyl, or 1-methylethyl; R²⁰ isselected from halo, methoxy, and methylsulfonyl; R²¹ is halo (morepreferably chloro); p is 0, 1, 2 or 3; and q is 0, 1, or 2.

More preferably, in compounds of formula (II), T is

or

Further preferred compounds are those of Formula (II), above, wherein Eand T are as immediately defined above; R²⁰ and the phenyl group towhich each R²⁰ is attached together form phenyl, 3-methylsulfonylphenyl,4-methylphenyl, or 3,4,5-trimethoxyphenyl, and R²¹ and the phenyl groupto which each R²¹ is attached together form 4-chlorophenyl or3,4-dichlorophenyl.

Utility

The compounds of the invention are CCR-3 receptor antagonists andinhibit eosinophil recruitment by CCR-3 chemokines such as RANTES,eotaxin, MCP-2, MCP-3 and MCP-4. Compounds of this invention andcompositions containing them are useful in the treatment ofeosiniphil-induced diseases such as inflammatory or allergic diseasesand including respiratory allergic diseases such as asthma, allergicrhinitis, hypersensitivity lung diseases, hypersensitivity pneumonitis,eosinophilic pneumonias (e.g., chronic eosinophilic pneumonia);inflammatory bowel diseases (e.g., Crohn's disease and ulcerativecolitis); and psoriasis and inflammatory dermatoses such as dermatitisand eczema.

Administration and Pharmaceutical Composition

In general, the compounds of this invention can be administered in atherapeutically effective amount by any of the accepted modes ofadministration for agents that serve similar utilities. The actualamount of the compound of this invention, i.e., the active ingredient,will depend upon numerous factors such as the severity of the disease tobe treated, the age and relative health of the subject, the potency ofthe compound used, the route and form of administration, and otherfactors.

Therapeutically effective amounts of compounds of Formula (I) may rangefrom approximately 0.01-20 mg per kilogram body weight of the recipientper day; preferably about 0.1-10 mg/kg/day. Thus, for administration toa 70 kg person, the dosage range would most preferably be about 7 mg to0.7 g per day.

In general, compounds of this invention will be administered aspharmaceutical compositions by any one of the following routes: oral,transdermal, inhalation (e.g., intranasal or oral inhalation) orparenteral (e.g., intramuscular, intravenous or subcutaneous)administration. A preferred manner of administration is oral using aconvenient daily dosage regimen which can be adjusted according to thedegree of affliction. Compositions can take the form of tablets, pills,capsules, semisolids, powders, sustained release formulations,solutions, suspensions, liposomes, elixirs, or any other appropriatecompositions. Another preferred manner for administering compounds ofthis invention is inhalation. This is an effective means for deliveringa therapeutic agent directly to the respiratory tract for the treatmentof diseases such as asthma and other similar or related respiratorytract disorders (see U.S. Pat. No. 5,607,915).

The choice of formulation depends on various factors such as the mode ofdrug administration and the bioavailability of the drug substance. Fordelivery via inhalation the compound can be formulated as liquidsolutions or suspensions, aerosol propellants or dry powder and loadedinto a suitable dispenser for administration. There are three types ofpharmaceutical inhalation devices—nebulizer inhalers, metered-doseinhalers (MDI) and dry powder inhalers (DPI). Nebulizer devices producea stream of high velocity air that causes the therapeutic agents (whichhas been formulated in a liquid form) to spray as a mist which iscarried into the patient's respiratory tract. MDI's typically have theformulation packaged with a compressed gas. Upon actuation, the devicedischarges a measured amount of therapeutic agent by compressed gas,thus affording a reliable method of administering a set amount of agent.DPI's administer therapeutic agents in the form of a free flowing powderthat can be dispersed in the patient's inspiratory air-stream duringbreathing by the device. In order to achieve a free flowing powder, thetherapeutic agent is formulated with an excipient, such as lactose. Ameasured amount of the therapeutic is stored in a capsule form and isdispensed to the patient with each actuation. Recently, pharmaceuticalformulations have been developed especially for drugs that show poorbioavailability based upon the principle that bioavailability can beincreased by increasing the surface area i.e., decreasing particle size.For example, U.S. Pat. No. 4,107,288 describes a pharmaceuticalformulation having particles in the size range from 10 to 1,000 nm inwhich the active material is supported on a crosslinked matrix ofmacromolecules. U.S. Pat. No. 5,145,684 describes the production of apharmaceutical formulation in which the drug substance is pulverized tonanoparticles (average particle size of 400 nm) in the presence of asurface modifier and then dispersed in a liquid medium to give apharmaceutical formulation that exhibits remarkably highbioavailability.

The compositions are comprised of in general, a compound of Formula (I)in combination with at least one pharmaceutically acceptable excipient.Acceptable excipients are non-toxic, aid administration, and do notadversely affect the therapeutic benefit of the compound of Formula (1).Such excipient may be any solid, liquid, semi-solid or, in the case ofan aerosol composition, gaseous excipient that is generally available toone of skill in the art.

Solid pharmaceutical excipients include starch, cellulose, talc,glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silicagel, magnesium stearate, sodium stearate, glycerol monostearate, sodiumchloride, dried skim milk and the like. Liquid and semi-solid excipientsmay be selected from glycerol, propylene glycol, water, ethanol andvarious oils, including those of petroleum, animal, vegetable orsynthetic origin, e.g., peanut oil, soybean oil, mineral oil, sesameoil, etc. Preferred liquid carriers, particularly for injectablesolutions, include water, saline, aqueous dextrose, and glycols.

Compressed gases may be used to disperse a compound of this invention inaerosol form. Inert gases suitable for this purpose are nitrogen, carbondioxide, etc.

For liposomal formulations of the drug for parenteral or oral deliverythe drug and the lipids are dissolved in a suitable organic solvent e.g.tert-butanol, cyclohexane (1% ethanol). The solution is lyopholized andthe lipid mixture is suspended in an aqueous buffer and allowed to forma liposome. If necessary, the liposome size can be reduced bysonication. (See Frank Szoka, Jr. and Demetrios Papahadjopoulos,“Comparative Properties and Methods of Preparation of Lipid Vesicles(Liposomes)”, Ann. Rev. Biophys. Bioeng., 9:467-508 (1980), and D. D.Lasic, “Novel Applications of Liposomes”, Trends in Biotech.,16:467-608, (1998)).

Other suitable pharmaceutical excipients and their formulations aredescribed in Remington's Pharmaceutical Sciences, edited by E. W. Martin(Mack Publishing Co., 18th ed., 1990).

The level of the compound in a formulation can vary within the fullrange employed by those skilled in the art. Typically, the formulationwill contain, on a weight percent (wt %) basis, from about 0.01-99.99 wt% of a compound of Formula (I) based on the total formulation, with thebalance being one or more suitable pharmaceutical excipients.Preferably, the compound is present at a level of about 1-80 wt %.Representative pharmaceutical formulations containing a compound ofFormula (I) are described below.

Testing

The CCR-3 antagonistic activity of the compounds of this invention canbe measured by in vitro assays such as ligand binding and chemotaxisassays as described in more detail below. In vivo activity can beassayed in the Ovalbumin induced Asthma in Balb/c Mice Model asdescribed in more detail below.

Abbreviations

For ease of reference, the following abbreviations are used in theSchemes and Examples below:

-   MeOH=methanol-   EtOH=ethanol-   EtOAc=ethyl acetate-   HOAc=acetic acid-   DCE=1,2-dichloroethane-   DMF=dimethylformamide-   PCC=pyridinium chlorochromate-   PDC=pyridinium dichromate-   TEA or Et₃N=triethylamine-   THF=tetrahydrofuran-   TFA=trifluoroacetic acid-   rt=room temperature    General Synthetic Schemes

The compounds of the present invention can be prepared in a number ofways known to one skilled in the art. Preferred methods include, but arenot limited to, the general synthetic procedures described below.

The starting materials and reagents used are either available fromcommercial suppliers such as Aldrich Chemical Co. (Milwaukee, Wis.,USA), Bachem (Torrance, Calif., USA), Enika Chemie or Sigma (St. Louis,Mo., USA), Maybridge (Dist: Ryan Scientific, P.O. Box 6496, Columbia,S.C. 92960), etc.; or are prepared by methods known to those skilled inthe art following procedures set forth in the literature such as Fieserand Fieser's Reagents for Organic Synthesis, Volumes 1-17 (John Wileyand Sons, 1991); Rodd's Chemistry of Carbon Compounds, Volumes 1-5 andSupplements (Elsevier Science Publishers, 1989), Organic Reactions,Volumes 1-40 (John Wiley and Sons, 1991), March's Advanced OrganicChemistry, (John Wiley and Sons, 1992), and Larock's ComprehensiveOrganic Transformations (VCH Publishers Inc., 1989). These schemes aremerely illustrative and various modifications to these schemes can bemade and will be suggested to one skilled in the art having referred tothis disclosure.

The starting materials and the intermediates of the reaction may beisolated and purified if desired using conventional techniques,including but not limited to filtration, distillation, crystallization,chromatography, and the like. Such materials may be characterized usingconventional means, including physical constants and spectral data. Inthe Schemes, the variables E, Q, Ar, Ar¹, R⁴, R²⁰, R²¹, p, q, etc., aredefined as set forth in the claims.

Step (a):

2-Methoxy-4-nitrobenzoic acid (1) in CH₂Cl₂ (97%) is reacted with BBr₃to provide the intermediate 2-hydroxy-4-nitro-benzoic acid, which upontreatment with SOCl₂ in solvent such as MeOH (95%), followed byhydrogenation and treatment with 10% Pd/C in EtOAc (98%) yields4-amino-2-hydroxy-benzoic acid methyl ester (2).

Step (b):

Hydrogenation of methyl ester (2) [e.g., 53 psi, 5% Rh on Al in aceticacid (70 h), 58° C.] affords 4-amino-2-hydroxy-cyclohexanecarboxylicacid methyl ester (3) as the acetic acid salt.

Step (c):

Reaction of compound (3) with mesitylene at elevated temperature (e.g.,165° C.) followed by cooling to rt yields5-hydroxy-2-aza-bicyclo[2.2.2]octan-3-one (4).

Step (d):

Compound (4) can be reacted with LAH in solvent (such as THF), withreflux under nitrogen to yield 2-aza-bicyclo[2.2.2]octane-5-ol (5).

Step (e):

Compound (5) can be N-Boc protected upon mixing with (Boc)₂O (86%) in anappropriate solvent (e.g., MeOH, TEA). N-Boc protected compound (5) canbe converted to N-Boc protected 2-aza-bicyclo[2.2.2]octan-5-one (6) upontreatment with PDC in an appropriate solvent such as CH₂Cl₂ (96%) orDMF.

Step (f):

N-Boc protected 2-aza-bicyclo[2.2.2]octan-5-one (6) can be converted tothe appropriately-substituted compound (7) via addition of3,4-di-ClPhCH₂P(O)(OEt)₂ in solvent such as THF at reduced temperature(e.g., 0° C.), followed by hydrogenation in solvent (such as EtOH:EtOAcor EtOAc), in the presence of PtO₂ (88%) and addition of TFA (99%).

Step (g):

Bicyclo-octane (7) can be reacted with an appropriately-substitutedaldehyde (8) in solvent such as CH₂Cl₂ (36%) or DCE upon addition ofMe₄NB(OAc)₃H or NaBH(OAc)₃, to yield compounds having the formula (Ia).

Step (a):

8-Methyl-8-aza-bicyclo[3.2.1]octan-3-one can be treated with 1-Cl-ethylchloroformate in an appropriate solvent such as MeOH (88%) or DCE,followed by addition of (Boc)₂O (98%) to afford compound (10).

Step (b):

Compound (10) can be treated with an appropriate phosphonic acid diethylester [e.g., Ar¹-Q-P(O)(MeO)₂], in the presence of base (e.g., potassiumt-amylate or NaH), and then hydrogenated in the presence of PtO₂ underH₂ and extracted with EtOAc to afford compound (11).

Steps (c)-(d):

Compound (11) can be treated with TFA in an appropriate solvent such asCH₂Cl₂, then reacted with (Boc)NHCH(R⁴)CO₂H, EDCI and/or HOBT in CH₂Cl₂,then reacted with TFA and refluxed with BH₃-THF at elevated temperature,followed by cooling and addietion of HCl (aq.) to provide compound (13).

Step (e):

Compound (13) can be treated with 4-toluoyl chloride and Et₃N in CH₂Cl₂or reacted with an appropriately-substituted isocyanate to affordcompounds of formulae (Ib) and/or (Ic).

The following Examples are provided as additional guidance for those ofskill in the art, and are not intended to limit the invention in anyway.

EXAMPLE 1

Step 1:

To a solution of 2-methoxy-4-nitro-benzoic acid (20 g, 0.102 mol) in 80ml of CH₂Cl₂ at 0° C. was added BBr₃ (1.0 M, 150 ml, 1.5 eq.). Theresulting mixture was allowed to warm to rt and stirred for 2 h. MeOHwas then added dropwise to quench the reaction at 0° C. and the volatilefraction was removed in vacuo. The residue was purified on a silica gelcolumn with 6/4/0.3 of hexane, EtOAc and HOAc to give 18 g of2-hydroxy-4-nitro-benzoic acid (M⁺: 183).

Step 2:

To a MeOH solution (200 ml) of 2-hydroxy-4-nitro-benzoic acid (18 g) wasadded concentrated sulfuric acid. The resulting mixture was heated toreflux for 24 h. After it was cooled to rt, volatile was removed invacuo. The residue was then partitioned between water and EtOAc. TheEtOAc layer was washed with water (2×), NaHCO₃ (sat.), NaCl (sat.),dried over Na₂SO₄ and concentrated. The crude product was purified on asilica gel column with 20% EtOAc in hexane to give 14.5 g of2-hydroxy-4-nitro-benzoic acid methyl ester as a solid (M⁺: 197).

Step 3:

2-Hydroxy-4-nitro-benzoic acid methyl ester (14 g) was dissolved in EtOH(100 ml) and THF (10 ml) and stirred under hydrogen (1 atm) in thepresence of PtO₂ for 24 h. The reaction mixture was then filteredthrough a celite bed. The filtrate was concentrated to give 12 g of4-Amino-2-hydroxy-benzoic acid methyl ester as a light yellow solid.

Step 4:

4-Amino-2-hydroxy-benzoic acid methyl ester (10 g) in 200 ml of HOAc washydrogenated under 56 psi pressure at 60° C. for 42 h. The volatile wasthen removed. The residue was stirred in 50 ml of 4/1 of ether/EtOH andprecipitate formed. Filtration gave 6.5 g of4-amino-2-hydroxy-cyclohexanecarboxylic acid methyl ester acetic acidsalt (m.p. 125.5-126.4° C.; M⁺: 173).

Step 5:

4-Amino-2-hydroxy-cyclohexanecarboxylic acid methyl ester acetic acidsalt (6.6 g) was heated to reflux in mesitylene (60 ml) for 3 h. Aftercooling to rt, solvent was decanted and the crystals were washed withhexane three times to provide 2.9 g of5-hydroxy-2-aza-bicyclo[2.2.2]octan-3-one (M⁺: 141).

Step 6:

To a solution of 5-hydroxy-2-aza-bicyclo[2.2.2]octan-3-one (2.6 g, 18mmol) in 60 ml of anhydrous THF was added 55 ml of LAH (1.0 M in THF).After the mixture was refluxed under N₂ for 24 h, it was cooled to rtand quenched with 2 ml of water, followed by 2 ml of 15% NaOH and 6 mlof water. The suspension was then stirred with MgSO₄ and filtered. Thefiltrate was acidified with 1.0 M of HCl in ether and then concentratedto afford 3.2 g of 2-aza-bicyclo[2.2.2]octan-5-ol as the HCl salt.

Step 7:

2-Aza-bicyclo[2.2.2]octan-5-ol (3.2 g, 0.02 mol) was mixed withdi-t-butyl dicarbonate (8.8 g, 2 eq.) in 60 ml of EtOH and 7.6 ml of TEA(3 eq.) and the resulting mixture was heated under N₂ at 60° C. for 3 h.The volatile fraction was removed and the residue was partitionedbetween EtOAc and water. The organic layer was washed with saturatedNaCl (aq.) and dried over Na₂SO₄ to afford N-Boc protected2-aza-bicyclo[2.2.2]octan-5-ol (3 g) (M⁺: 227).

Step 8:

To a solution of N-Boc protected 2-aza-bicyclo[2.2.2]octan-5-ol (3.0 g,31.4 mmol) in DMF at 0° C. was added 30 g of PDC (7 eq.). The resultingmixture was stirred for 6 h and filtered through a celite bed. Thefiltrate was concentrated and purified on a silica gel column with 20%EtOAc in hexane to give 1.74 g of N-Boc protected2-aza-bicyclo[2.2.2]octan-5-one (M⁺: 225).

Step 9:

Triethylphosphite (4.28 ml) was added dropwise to3,4-dichlorobenzylbromide (6.0 g, 25 mmol) at rt with stirring. After 1ml of triethylphosphite was added, the mixture was heated to 80° C.until an exothermic reaction was started. The remaining phosphite wasadded at a rate sufficient to maintain the reflux. After the additionwas completed, the mixture was heated to reflux for 1 h and allowed tocool. The crude product was distilled under vacuum at 155-158° C./1-2torr to afford (3,4-dichloro-benzyl)-phosphonic acid diethyl ester (6.5g).

Step 10:

To a suspension of NaH (53 mg, 2.1 mmol) in 8 ml of THF with 13 mg of15-crown-5 (3%) was added N-Boc protected2-aza-bicyclo[2.2.2]octan-5-one (0.45 g, 2 mmol) (from Step 8) and(3,4-Dichloro-benzyl)-phosphonic acid diethyl ester (0.594, 2 mmol)(from Step 9) in 4 ml of THF dropwise at 0° C. under N₂. Hydrogenevolution was observed and the solution turned yellow. After thecompletion of the addition, the mixture was stirred at 0° C. for 1 h andat rt for 2 h. It was then quenched with water and extracted with EtOAc.The EtOAc layer was washed with NaCl (sat.), dried over Na₂SO₄ andconcentrated. Purification on a silica gel column with 20% EtOAc inhexane gave 0.45 g of5-(3,4-dichlorobenzylidene)-2-aza-bicyclo[2.2.2]octane-2-carboxylic acidtert-butyl ester (M⁺: 367).

Step 11:

5-(3,4-Dichloro-benzylidene)-2-aza-bicyclo[2.2.2]octane-2-carboxylicacid tert-butyl ester (0.45 g) was stirred under 1 atm of H₂ in 20 ml of1:1 of EtOH:EtOAc in the presence of PtO₂ for 20 min. The reactionmixture was filtered through celite and concentrated to give 0.4 g of5-(3,4-dichloro-benzyl)-2-aza-bicyclo[2.2.2]octane-2-carboxylic acidtert-butyl ester (M⁺: 369).

Step 12:

To a solution of5-(3,4-dichloro-benzyl)-2-aza-bicyclo[2.2.2]octane-2-carboxylic acidtert-butyl ester (0.24 g, 0.65 mmol) in 2 mL of CH₂Cl₂ was added 1 mL ofTFA. After the mixture was stirred at rt for 1 h, it was quenched withNaHCO₃ (sat.). It was then extracted with CH₂Cl₂ and the organic layerwas washed with NaCl (sat.), dried over Na₂SO₄ and concentrated to give170 mg of 5-(3,4-Dichloro-benzyl)-2-aza-bicyclo[2.2.2]octane (M⁺+1:270).

Step 13:

To a solution of valinol (DL, 5.0 g, 0.048 mol) in 200 mL of CH₂Cl₂ inthe presence of Et₃N (18.4 mL, 3 eq.) at 0° C. was added4-methyl-benzoyl chloride (7.1 mL, 1.1 eq.) in 50 mL of CH₂Cl₂ dropwise.After the addition was complete, the mixture was stirred at rtovernight. It was then quenched with water, and the organic layer wasextracted with CH₂Cl₂, washed with NaCl (sat.), dried over Na₂SO₄ andconcentrated. Column purification with 2/2/6 of acetone/CH₂Cl₂/hexanegave 7.8 g of white solidN-(1-hydroxymethyl-2-methyl-propyl)-4-methyl-benzamide (M⁺: 221).

Step 14:

To a suspension of PCC in 20 mL of CH₂Cl₂ was addedN-(1-hydroxymethyl-2-methyl-propyl)-4-methyl-benzamide (2.2 g, 10 mmol)in 15 mL of CH₂Cl₂. After the mixture was stirred for 1.5 h, it wasdiluted with Et₂O and filtered through a celite bed. The filtrate wasconcentrated and the residue purified on a silica gel column with 20%EtOAc in hexane to give 0.75 g ofN-(1-formyl-2-methyl-propyl)-4-methyl-benzamide as a white solid (M⁺+1:220).

Step 15:

To a solution of N-(1-formyl-2-methyl-propyl)-4-methyl-benzamide (0.083g, 1.2 eq.) and 5-(3,4-dichloro-benzyl)-2-aza-bicyclo[2.2.2]octane(0.085 g, 0.32 mmol) in 2 mL of CH₂Cl₂ at rt was added NaBH(OAc)₃ (96mg, 1.1 eq.). After the resulting mixture was stirred for 16 h, it wasquenched with NaHCO₃ (sat.) and extracted with EtOAc. The organic layerwas washed with NaCl (sat.), dried over Na₂SO₄ and concentrated.Preparative TLC with 5% MeOH in CH₂Cl₂ and recrystallization from CH₂Cl₂and hexane gave 50 mg of the compound of Example 1,N-(1-[5-(3,4-dichlorobenzyl)-2-aza-bicyclo[2.2.2]oct-2-ylmethyl]2-methylpropyl)-4-methylbenzamide,(M⁺+1: 473).

EXAMPLE 24-[5-(3,4-Dichloro-benzyl)-2-aza-bicyclo[2.2.2]oct-2-ylmethyl]-1,5-dimethyl-2-phenyl-1,2-dihydro-pyrazol-3-one

To a solution of 5-(3,4-dichloro-benzyl)-2-aza-bicyclo[2.2.2]octane(0.05 g, 0.35 mmol) in 2 ml of DCE was added1,5-dimethyl-3-oxo-2-phenyl-2,3-dihydro-1H-pyrazole-4-carbaldehyde (90mg, 1.2 eq.), and the mixture was stirred at rt for 15 min. NaBH(OAc)₃(0.11 g, 1.5 eq.) was then added and the mixture was stirred at rtovernight, then with NaHCO₃ (sat.), and extracted with EtOAc. The EtOAclayer was washed with NaCl (sat.), dried over Na₂SO₄ and concentrated.Preparative TLC with 10% (10% NH₄OH in MeOH) in CH₂Cl₂ gave 82 mg ofExample 2 (M⁺+1: 470).

EXAMPLE 3N-{1-[3-(3,4-Dichloro-benzyl)-8-aza-bicyclo[3.2.1]oct-8-ylmethyl]-2-methyl-propyl}-4-methyl-benzamide

Step 1:

To a solution of 8-methyl-8-aza-bicyclo[3.2.1]octan-3-one (6.8 g, 0.05mol) in 80 mL of DCE was added 1-chloroethyl chloroformate (8 mL, 1.5eq.) dropwise at 0° C. After the addition was complete, the reactionmixture was allowed to warm to rt and was then heated to reflux for 3 h.After volatile was removed, the solid residue was dissolved in 100 mL ofMeOH and then heated to reflux for 45 min. The volatile fraction wasthen removed again in vacuo and 8-aza-bicyclo[3.2.1]octan-3-onehydrochloride was recrystallized from MeOH/Et₂O as a solid product (58%yield) (4.58 g).

Step 2:

To a solution of 8-aza-bicyclo[3.2.1]octan-3-one hydrochloride (4.5 g,0.028 mol) in 100 mL of EtOH was added carbonic acid di-tert-butyl ester(12 g, 2 eq.) and 11 mL of TEA. The resulting mixture was heated at 60°C. for 3 h. The volatile fraction was removed and the residue waspartitioned between EtOAc and water. The EtOAc layer was washed withsaturated sodium chloride, dried over Na₂SO₄ and concentrated. Silicagel column purification with 20% EtOAc in hexane gave3-oxo-8-aza-bicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester(6.25 g).

Step 3:

To a suspension of NaH (0.24 g, 1.2 eq.) and 15-crown-5 (88 mg, 5%) in32 mL of THF at 0° C. was added a solution of3-oxo-8-aza-bicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester (1.8g, 8 mmol) and (3,4-dichloro-benzyl)-phosphonic acid diethyl ester (2.6g, 8.8 mmol) in 16 mL of THF. The resulting mixture was stirred at 0° C.for 1 h and at rt for 5 h. It was then quenched with H₂O and extractedwith EtOAc. The organic layer was separated, washed with NaCl (sat.) anddried over Na₂SO₄. Column purification with 20% EtOAc in hexane provided1.2 g of the starting material(3-Oxo-8-aza-bicyclo[3.2.1]octane-8-carboxylic acid tert-butyl ester)and 1 g of the desired product,3-(3,4-dichloro-benzylidene)-8-aza-bicyclo[3.2.1]octane-8-carboxylicacid tert-butyl ester (M⁺+1: 368).

Step 4:

3-(3,4-Dichloro-benzylidene)-8-aza-bicyclo[3.2.1]octane-8-carboxylicacid tert-butyl ester (1.0 g, 2.7 mmol) in 10 mL of EtOH and 10 mL ofEtOAc was stirred with 25 mg of PtO₂ under 1 atm of H₂ at rt for 3 h. Itwas then filtered through a celite bed and the filtrate wasconcentrated. The residue was purified on a silica gel column to give0.84 g of3-(3,4-dichloro-benzyl)-8-aza-bicyclo[3.2.1]octane-8-carboxylic acidtert-butyl ester (M⁺+1: 370).

Step 5:

3-(3,4-Dichloro-benzyl)-8-aza-bicyclo[3.2.1]octane-8-carboxylic acidtert-butyl ester (0.35 g, 0.94 mmol) was stirred in 2 mL of CH₂Cl₂ with0.5 mL of TFA at rt for 3 h. After being quenched with 20% NaOH to basicmedium, the mixture was extracted with EtOAc. The organic layer waswashed with NaCl (sat.), dried over Na₂SO₄ and concentrated to give 0.25g of 3-(3,4-Dichloro-benzyl)-8-aza-bicyclo[3.2.1]octane.

Step 6:

To a solution of N-(1-formyl-2-methyl-propyl)-4-methyl-benzamide (0.11g, 0.44 mmol) (prepared according to example 1, step 13 and 14) and3-(3,4-dichloro-benzyl)-8-aza-bicyclo[3.2.1]octane (0.11 g, 0.4 mmol) in2 mL of CH₂Cl₂ at rt was added NaBH(OAc)₃. After the resulting mixturewas stirred for 12 h, it was quenched with NaHCO₃ (sat.) and extractedwith EtOAc. The organic layer was washed with NaCl (sat.), dried overNa₂SO₄ and concentrated. Preparative TLC with 5% (10% NH₄OH in MeOH) inCH₂Cl₂ and acidification with HCl gave 125 mg of Example 3 (m.p.230-235° C.; M⁺+1: 473)

EXAMPLE 4(R)-1-{2-[4-(4-Chloro-benzyl)-2-ethyl-6-methyl-piperidin-1-yl]-1-methyl-ethyl}-3-(3,4,5-trimethoxy-phenyl)-urea

Step 1:

To a stirred solution of trimethylphosphite (0.57 ml, 4.9 mmol) wasadded 4-chlorobenzylbromide (1.0 g, 4.9 mmol) at rt. The resultingmixture was stirred at rt for 5 min. and then heated in an oil bath at80° C. for 20 min. It was cooled to rt and the product purified on asilica-gel column with 25% EtOAc in hexane to give 1.05 g of(4-chloro-benzyl)-phosphonic acid dimethyl ester (93%).

Step 2:

3-(4-Chloro-benzylidene)-8-aza-bicyclo[3.2.1]octane-8-carboxylic acidtert-butyl ester was prepared according to the procedure described inexample 3, step 3, but substituting (3,4-dichloro-benzyl)-phosphonicacid diethyl ester for (4-chloro-benzyl)-phosphonic acid dimethyl ester.Hydrogenation of3-(4-chloro-benzylidene)-8-aza-bicyclo[3.2.1]octane-8-carboxylic acidtert-butyl ester under atmospheric hydrogen in the presence of PtO₂ inEtOH gave 3-(4-chloro-benzyl)-8-aza-bicyclo[3.2.1]octane-8-carboxylicacid tert-butyl ester in 65% yield (M⁺: 335).

Step 3:

3-(4-chloro-benzyl)-8-aza-bicyclo[3.2.1]octane-8-carboxylic acidtert-butyl ester (0.88 g, 2.6 mmol) was stirred in 10 mL of CH₂Cl₂ with10 mL of TFA at rt for 45 min. After it was quenched with 20% NaOH tobasic medium, the mixture was extracted with EtOAc. The organic layerwas washed with NaCl (sat.), dried over Na₂SO₄ and concentrated to give0.52 g of 3-(4-Dichloro-benzyl)-8-aza-bicyclo[3.2.1]octane (85%).

Step 4:

A mixture of 3-(4-Dichloro-benzyl)-8-aza-bicyclo[3.2.1]octane,(D)-2-t-butoxy-carbonylamino-propionic acid, HOBT and EDCI in CH₂Cl₂ wasstirred at rt overnight. It was then quenched with Na₂CO₃ (sat.) andextracted with CH₂Cl₂ The organic layer was washed with NaCl (sat.) anddried over Na₂SO₄. The crude product({2-[3-(4-chloro-benzyl)-8-aza-bicyclo[3.2.1]oct-8-yl]-1-methyl-2-oxo-ethyl}carbamicacid tert-butyl ester) was purified on a silica-gel column with 25%acetone in CH₂Cl₂. It was then reacted with TFA in CH₂Cl₂ at rt for 2 h.The work-up procedure described in step 3, above, gave the desiredproduct, which was dissolved in THF and heated to reflux with BH₃-THFfor 4 h. After cooling to rt, the reaction flask was further cooled to0° C. and 6N HCl was added. The resulting mixture was heated again toreflux for 1 h, cooled to rt and quenched with Na₂CO₃ (sat.). Themixture was extracted with EtOAc and the organic layer was washed withNaCl (sat.) and dried over Na₂SO₄. The crude product was purified on asilica-gel with 40% EtOAc in hexane and followed by 1% iPr-NH₂, 9% MeOHin EtOAc to give2-[3-(4-chlorobenzyl)-8-aza-bicyclo[3.2.1]oct-8-yl]-1-methyl-ethylaminein 55% yield.

Step 5:

To a solution of2-[3-(4-chlorobenzyl)-8-aza-bicyclo[3.2.1]oct-8-yl]-1-methyl-ethylaminein CH₂Cl₂ was added 3,4,5-trimethoxyphenylisocyanate at −78° C. Afterthe addition, it was allowed to warm up to rt where it was stirred for 4h. It was then diluted with CH₂Cl₂. The organic layer was washed withNaCl (sat.) and dried over Na₂SO₄. The crude product was purified on asilica-gel column with 3% (10% iPr-NH₂ in MeOH) in EtOAc to give 55% ofExample 4 (m.p. 115-120° C., M⁺: 502).

EXAMPLE 51-{2-[4-(4-Chloro-benzyl)-2-ethyl-6-methyl-piperidin-1-yl]-1-methyl-ethyl}-3-(3-methanesulfonylphenyl)urea

To a solution of triphosgene (0.22 g, 0.33 eq.) in CH₂Cl₂ was added3-methane-sulfonyl-phenylamine hydrochloride (0.5 g, 2.4 mmol), followedby the dropwise addition of TEA (0.37 ml, 1.1 eq.). The mixture washeated to 40° C. for 30 min. It was allowed to cool to rt and stirredfor an additional 45 min. The isocyanate solution was then added to asolution of2-[3-(4-chlorobenzyl)-8-aza-bicyclo[3.2.1]oct-8-yl]-1-methyl-ethylamine(0.18 g, 0.6 mmol) (prepared according to the procedure described inexample 4, steps 1 to 4) in 6 ml of CH₂Cl₂. The mixture was stirredovernight at rt. It was diluted with CH₂Cl₂ and the organic layer waswashed with Na₂CO₃ (sat.) twice, NaCl (sat.) and dried over Na₂SO₄ Thecrude mixture was purified on a silica-gel column with 1% iPrNH₂, 9%MeOH in EtOAc to give 55% of Example 5 (M⁺: 490).

EXAMPLES 6-10

Compounds described below in Table 1 were prepared following the same orsimilar methods described above for Examples 1-5. TABLE 1 Ex. No.Structure Compound Name Data 6

4-[3-(3,4-dichloro- benzyl)-8-aza- bicyclo[3.2.1]oct-8-ylmethyl]-1,5-dimethyl- 2-phenyl-1,2-dihydro- pyrazol-3-one M⁺ + 1:470 7

1-{1-[4-(4-Chloro- benzyl)-2-ethyl-6- methyl-piperidin-1-yl-methyl]-2-hydroxy- propyl}-3-(3,4,5-tri- methoxyphenyl)-urea m.p.118-123° C.; M⁺: 532 8

1-{1-[4-(4-Chloro- benzyl)-2-ethyl-6- methyl-piperidin-1-yl-methyl]-2-hydroxy- propyl}-3-(3-methane- sulfonyl-phenyl)-urea m.p.94-103° C.; M⁺: 520. 9

1-{2-[4-(4-chloro- benzyl)-2-ethyl-6- methyl-piperidin-1-yl]-ethyl}-3-(3,4,5-trimeth- oxy-phenyl)-urea m.p. 107-111° C.; M⁺: 488. 10

1-{2-[4-(4-Chloro- benzyl)-2-ethyl-6- methyl-piperidin-1-yl]-ethyl}-3-(3-methane- sulfonyl-phenyl)-urea M⁺:476.

EXAMPLE 11 Formulation Examples

The following are representative pharmaceutical formulations containinga compound of Formula (I).

Tablet Formulation

The following ingredients are mixed intimately and pressed into singlescored tablets. Quantity per Ingredient tablet, mg compound of thisinvention 400  cornstarch 50 croscarmellose sodium 25 lactose 120 magnesium stearate  5Capsule Formulation

The following ingredients are mixed intimately and loaded into ahard-shell gelatin capsule. Quantity per Ingredient capsule, mg compoundof this invention 200 lactose, spray-dried 148 magnesium stearate  2Suspension Formulation

The following ingredients are mixed to form a suspension for oraladministration. Ingredient Amount compound of this invention 1.0 gfumaric acid 0.5 g sodium chloride 2.0 g methyl paraben 0.15 g propylparaben 0.05 g granulated sugar 25.5 g sorbit (70% solution) 12.85 gVeegum K (Vanderbilt Co.) 1.0 g flavoring 0.035 ml colorings 0.5 mgdistilled water q.s. to 100 mlInjectable Formulation

The following ingredients are mixed to form an injectable formulation.Ingredient Amount compound of this invention 0.2 g sodium acetate buffersolution, 0.4 M 2.0 ml HCl (1 N) or NaOH (1 N) q.s. to suitable pH water(distilled, sterile) q.s. to 20 mlLiposomal Formulation

The following ingredients are mixed to form a liposomal formulation.Ingredient Amount compound of this invention 10 mgL-.alpha.-phosphatidylcholine 150 mg tert-butanol 4 ml

Freeze dry the sample and lyophylize overnight. Reconstitute the samplewith 1 ml 0.9% saline solution. Liposome size can be reduced bysonication.

EXAMPLE 12

CCR-3 Receptor Binding Assay-In Vitro

The CCR-3 antagonistic activity of the compounds of the invention wasdetermined by their ability to inhibit the binding of ¹²⁵I eotaxin toCCR-3 L1.2 transfectant cells (see Ponath, P. D. et al., J. Exp. Med.,183:2437-48, (1996)).

The assay was performed in Costar 96-well polypropylene round bottomplates. Test compounds were dissolved in DMSO and then diluted withbinding buffer (50 mM HEPES, 1 mM CaCl₂, 5 mM MgCl₂, 0.5% bovine serumalbumin (BSA), 0.02% sodium azide, pH 7.24) such that the final DMSOconcentration was 2%. 25 μl of the test solution or only buffer withDMSO (control samples) was added to each well, followed by the additionof 25 μl of ¹²⁵I-eotaxin (100 pmol) (NEX314, New England Nuclear,Boston, Mass.) and 1.5×10⁵ of the CCR-3 L1.2 transfected cells in 25 μlbinding buffer. The final reaction volume was 75 μl.

After incubating the reaction mixture for 1 hour at rt, the reaction wasterminated by filtering the reaction mixture through apolyethylenimine-treated Packard Unifilter GF/C filter plate (Packard,Chicago, Ill.). The filters were washed four times with ice cold washbuffer containing 10 mm HEPES and 0.5 M sodium chloride (pH 7.2) anddried at 65° C. for approximately 10 minutes. 25 μl/well ofMicroscint-20® scintillation fluid (Packard) was added and theradioactivity retained on the filters was determined by using thePackard TopCount®

Compounds of this invention were active in this assay. RepresentativeIC₅₀ values (nM) obtained with this binding assay for certain compoundslisted in Examples herein are shown below in Table 2. TABLE 2 Ex. No.IC₅₀ (nM) 2 331 4 11 6 965 7 8.2

EXAMPLE 13

Inhibition of Eotaxin Mediated Chemotaxis of CCR-3 L1.2 TransfectantCells—In Vitro Assay

The CCR-3 antagonistic activity of the compounds of this invention canbe determined by measuring the inhibition of eotaxin mediated chemotaxisof the CCR-3 L1.2 transfectant cells, using a slight modification of themethod described in Ponath, P. D. et al., J. Clin. Invest. 97:604-12(1996). The assay is performed in a 24-well chemotaxis plate (CostarCorp., Cambridge, Mass.). CCR-3 L1.2 transfectant cells are grown inculture medium containing RPMI 1640, 10% Hyclone® fetal calf serum, 55mM 2-mercaptoethanol and Geneticin 418 (0.8 mg/ml). 18-24 hours beforethe assay, the transfected cells are treated with n-butyric acid at afinal concentration of 5 mM/1×10⁶ cells/ml, isolated and resuspended at1×10⁷ cells/ml in assay medium containing equal parts of RPMI 1640 andMedium 199 (M 199) with 0.5% bovine serum albumin.

Human eotaxin suspended in phosphate buffered saline at 1 mg/ml is addedto bottom chamber in a final concentration of 100 nm. Transwell cultureinserts (Costar Corp., Cambridge, Mass.) having 3 micron pore size areinserted into each well and L1.2 cells (1×10⁶) are added to the topchamber in a final volume of 100 μl. Test compounds in DMSO are addedboth to the top and bottom chambers such that the final DMSOconcentration is 0.5%. The assay is performed against two sets ofcontrols. The positive control contained cells with no test compound inthe top chamber and only eotaxin in the lower chamber. The negativecontrol contains cells with no test compound in the top chamber andneither eotaxin nor test compound in lower chamber. The plate isincubated at 37° C. After 4 hours, the inserts are removed from thechambers and the cells that have migrated to the bottom chamber arecounted by pipetting out 500 μl of the cell suspension from the lowerchamber to 1.2 ml Cluster tubes (Costar) and counting them on a FACS for30 seconds.

EXAMPLE 14

Inhibition of Eotaxin Mediated Chemotaxis of Human Eosinophils—In VitroAssay

The ability of compounds of the invention to inhibit eotaxin mediatedchemotaxis of human eosinophils can be assessed using a slightmodification of procedure described in Carr, M. W. et al., Proc. Natl.Acad. Sci. USA, 91:3652-56 (1994). Experiments are performed using 24well chemotaxis plates (Costar Corp., Cambridge, Mass.). Eosinophils areisolated from blood using the procedure described in PCT ApplicationWO96/22371. The endothelial cells used are the endothelial cell line ECV304 obtained from European Collection of Animal Cell Cultures (PortonDown, Salisbury, U.K.). Endothelial cells are cultured on 6.5 mmdiameter Biocoat®. Transwell tissue culture inserts (Costar Corp.,Cambridge, Mass.) with a 3.0 μm pore size. Culture media for ECV 304cells consists of M199, 10% Fetal Calf Serum, L-glutamine andantibiotics. Assay media consists of equal parts RPMI 1640 and M199,with 0.5% BSA. 24 hours before the assay 2×10⁵ ECV 304 cells are platedon each insert of the 24-well chemotaxis plate and incubated at 37° C.20 nM of eotaxin diluted in assay medium is added to the bottom chamber.The final volume in bottom chamber is 600 μl. The endothelial coatedtissue culture inserts are inserted into each well. Eosinophil cells(10⁶) suspended in 100 μl assay buffer are added to the top chamber.Test compounds dissolved in DMSO are added to both top and bottomchambers such that the final DMSO volume in each well was 0.5%. Theassay is performed against two sets of controls. The positive controlcontains cells in the top chamber and eotaxin in the lower chamber. Thenegative control contains cells in the top chamber and only assay bufferin the lower chamber. The plates are incubated at 37° C. in 5% CO₂/95%air for 1-1.5 hours.

The cells that migrate to the bottom chamber are counted using flowcytometry. 500 μl of the cell suspension from the lower chamber areplaced in a tube, and relative cell counts are obtained by acquiringevents for a set time period of 30 seconds.

EXAMPLE 15 Inhibition of Eosinophil Influx into the Lungs of Ovalbumin

Sensitized Balb/c Mice by CCR-3 Antagonist—In Vivo Assay

The ability of the compounds of the invention to inhibit leukocyteinfiltration into the lungs can be determined by measuring theinhibition of eosinophil accumulation into the bronchioalveolar lavage(BAL) fluid of Ovalbumin (OA)-sensitized balb/c mice after antigenchallenge by aerosol. Briefly, male balb/c mice weighing 20-25 g aresensitized with OA (10 μg in 0.2 ml aluminum hydroxide solution)intraperitoneally on days 1 and 14. After a week, the mice are dividedinto ten groups. Test compound or only vehicle (control group) oranti-eotaxin antibody (positive control group) is administered eitherintraperitoneally, subcutaneously or orally. After 1 hour, the mice areplaced in a Plexiglass box and exposed to OA aerosol generated by aPARISTAR™ nebulizer (PARI, Richmond, Va.) for 20 minutes. Mice whichhave not been sensitized or challenged are included as a negativecontrol. After 24 or 72 hours, the mice are anesthetized (urethane,approx. 1 g/kg, i.p.), a tracheal cannula (PE 60 tubing) is inserted andthe lungs are lavaged four times with 0.3 ml PBS. The BAL fluid istransferred into plastic tubes and kept on ice. Total leukocytes in a 20μl aliquot of the BAL fluid is determined by Coulter Counter™. (Coulter,Miami, Fla.). Differential leukocyte counts are made on Cytospin™preparations which have been stained with a modified Wright's stain(DiffQuick™.) by light microscopy using standard morphological criteria.

1. A compound having the formula:

wherein T is

where R⁶ is taken together with one of R⁷ and R⁸ to form a bridge of oneto two carbon atoms, and the other of R⁷ and R⁸ is selected fromhydrogen and R⁹; Ar and Ar¹ are, independently of each other, aryl orheteroaryl, with the proviso that Ar and Ar¹ are not simultaneouslyphenyl; F is alkylene, alkenylene, or a bond; E is selected from—C(═O)N(R¹⁰)—, —SO₂N(R¹⁰)—, —N(R¹¹)C(═O)N(R¹⁰)—, —N(R¹¹)SO₂N(R¹⁰)—,—N(R¹¹)C(═S)N(R¹⁰)—, —N(R¹¹)C(═O)—, —N(R¹¹)SO₂—, —N(R¹²)C(═O)CH(R¹³)—,and CH(R¹³)C(═O)N(R¹²)—, where: R¹⁰, R¹¹, R¹², and R¹³ are,independently of each other, hydrogen, alkyl, acyl, haloalkyl,cycloalkyl, cycloalkylalkyl, aryl, aralkyl, aralkenyl, heteroaryl,heteroaralkyl, heterocycloalkyl, heteroalkyl, or -(alkylene)-C(═O)-Z,where Z is alkyl, haloalkyl, alkoxy, haloalkyloxy, hydroxy, amino, mono-or disubstituted amino, aryl, aralkyl, aryloxy, aralkyloxy, heteroaryl,heteroaryloxy, or heteroaralkyloxy; or alternatively, R¹² and R¹³ may betaken together with the nitrogen and carbon atoms to which they areattached, respectively, to form a heterocyclyl or heteroaryl ringoptionally substituted with up to two groups selected from R¹⁴; R³ andR⁴ are, independently of each other, hydrogen, alkyl, alkenyl,haloalkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heteroaryl,heteroaralkyl, heterocyclyl, heterocyclylalkyl, heteroalkyl,-(alkylene)-C(═O)-Z¹, or -(alkylene)-C(O)₂Z¹, where Z¹ is alkyl,haloalkyl, alkoxy, haloalkyloxy, hydroxy, amino, mono- or disubstitutedamino, aryl, aralkyl, aryloxy, aralkyloxy, heteroaryl, heteroaryloxy, orheteroaralkyloxy; R⁵ is hydrogen or alkyl; Q is —C(═O)— or C₁₋₂alkylene;R⁹ is attached to any available carbon atom of ring T and is selectedfrom lower alkyl, hydroxy, lower alkoxy, halo, cyano, trifluoromethyl,trifluoromethoxy, or a lower alkyl substituted with one of hydroxy,lower alkoxy, halo, cyano, trifluoromethyl, or trifluoromethoxy; R¹⁴ isselected from lower alkyl, hydroxy, lower alkoxy, halo, cyano,trifluoromethyl, trifluoromethoxy, and a lower alkyl substituted withone of hydroxy, lower alkoxy, halo, cyano, trifluoromethyl, ortrifluoromethoxy; m is 0 or 1; and n is 0 to 4; and or apharmaceutically acceptable salt thereof.
 2. A compound according toclaim 1, or a pharmaceutically acceptable salt thereof, wherein: Ar orAr¹ is optionally substituted phenyl; F is a bond; E is selected from—C(═O)N(R¹⁰)—, —N(R¹¹)C(═O)N(R¹⁰)—, —N(R¹¹)C(═O)—, —N(R¹²)C(═O)CH(R¹³)—,and CH(R¹³)C(═O)N(R¹²)—, where: R¹⁰, R¹¹, R¹², and R¹³ are,independently of each other, hydrogen or alkyl; or alternatively, R¹²and R¹³ may be taken together with the nitrogen and carbon atoms towhich they are attached, respectively, to form a heterocyclyl orheteroaryl ring optionally substituted with up to two groups selectedfrom R¹⁴; R³ and R⁴ are, independently of each other, hydrogen, alkyl,alkenyl, haloalkyl, heteroalkyl, or -(alkylene)-C(═O)-Z¹, where Z¹ isalkyl, haloalkyl, alkoxy, haloalkyloxy, hydroxy, amino, mono- ordisubstituted amino, aryl, aralkyl, aryloxy, aralkyloxy, heteroaryl,heteroaryloxy, or heteroaralkyloxy; Q is —CH₂—; R⁹ and R¹⁴ areindependently selected from methyl, ethyl, hydroxy, methoxy, halo,cyano, trifluoromethyl, or trifluoromethoxy; and n is 0 to
 2. 3. Acompound according to claim 1, or a pharmaceutically acceptable saltthereof, wherein T is selected from the group consisting of:

and R⁹ is attached to any available carbon atom of ring T and isselected from lower alkyl and hydroxy, and n is 0 to
 2. 4. A compound ofclaim 1, or a pharmaceutically acceptable salt thereof, wherein: Ar is aphenyl ring optionally substituted with one, two or three substituentsselected from alkyl, heteroalkyl, alkoxy, —COR¹⁵, —SO₂R¹⁷,methylenedioxy, hydroxy, halo, acylamino, amino, mono- or disubstitutedamino, —CONR¹⁵R¹⁶, -(alkylene)-CONR¹⁵R¹⁶, —COOR¹⁵, -(alkylene)-COOR¹⁵and —NR¹⁶SO₂R¹⁷; R¹⁵ and R¹⁶ are each independently hydrogen or alkyl;and R¹⁷ is alkyl, amino or mono or disubstituted amino.
 5. A compound ofclaim 4, or a pharmaceutically acceptable salt thereof, wherein Ar isselected from phenyl, 4-chlorophenyl, 4-methylphenyl, 4-methoxyphenyl,3-methylsulfonylphenyl, 3,5-dimethoxyphenyl, 3,4-dimethoxyphenyl, and3,4,5-trimethoxyphenyl.
 6. A compound of claim 1, or a pharmaceuticallyacceptable salt thereof, wherein F is a bond.
 7. A compound of claim 1,or a pharmaceutically acceptable salt thereof, wherein E is—C(═O)N(R¹⁰)—, —N(R¹⁰)C(═O)N(R¹¹)—, or N(R¹²)C(═O)CH(R¹³)—, where R¹⁰and R¹¹ are hydrogen or lower alkyl, and R¹² and R¹³ are taken togetherwith the nitrogen and carbon atoms to which they are attached,respectively, to form

where R¹⁸ and R¹⁹ are selected from hydrogen and lower alkyl.
 8. Acompound of claim 7, or a pharmaceutically acceptable salt thereof,wherein E is

and m is
 0. 9. A compound of claim 1, or a pharmaceutically acceptablesalt thereof, wherein R³ is hydrogen; and R⁴ is hydrogen, methyl, ethyl,1-methylethyl, isopropyl, 1-hydroxyethyl or 2-hydroxyethyl.
 10. Acompound of claim 1, or a pharmaceutically acceptable salt thereof,wherein R³ is hydrogen; and R⁴ is 1-methylethyl.
 11. A compound of claim1, or a pharmaceutically acceptable salt thereof, wherein T is


12. A compound of claim 1, or a pharmaceutically acceptable saltthereof, wherein Q is —CH₂—.
 13. A compound of claim 1, or apharmaceutically acceptable salt thereof, wherein: Ar¹ is a phenyl ringoptionally substituted with one, two or three substituent selected fromalkyl, heteroalkyl, alkoxy, halo, trifluoromethyl, nitro, or mono- ordisubstituted amino.
 14. A compound of claim 1, or a pharmaceuticallyacceptable salt thereof, wherein: Ar¹ is 4-chlorophenyl or3,4-dichlorophenyl. 15-17. (canceled)
 18. A pharmaceutical compositioncomprising a therapeutically effective amount of a compound of claim 1and a pharmaceutically acceptable excipient. 19-20. (canceled)